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Preparation method of indirect ELISA detection kit based on theileria equi EMA1

A technology for detecting kits and surface proteins, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of time-consuming, poor specificity, and expensive detection kits, and achieve the effect of eradicating harm and ensuring healthy development.

Inactive Publication Date: 2018-10-09
XINJIANG AGRI UNIV
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AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a method for preparing an indirect ELISA detection kit based on the surface protein of the elegans merozoites 1, to prepare highly efficient and specific surface proteins of the merozoites of the elegans merozoites for ELISA diagnosis, and to solve the problem of conventional detection Due to the disadvantages of expensive kits, poor specificity, and time-consuming, the establishment of rELISA detection method for equine worm disease is suitable for clinical diagnosis and epidemiological monitoring of equine worm disease

Method used

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Embodiment Construction

[0009] 1. Obtain local positive target gene fragments

[0010] (1) By comparing the conserved sequence of the EMA1 gene of Theileria equatus registered in GenBank, a pair of primers were designed using PrimerPrimier5 and PrimerExpress3.0 software. The target fragment size was 819bp, which was synthesized by Beijing Dingguo Biotechnology Co., Ltd.

[0011] Primer name Primername

Sequence (5'-3')sequence

EMA1-P1

5'-C GGATCC ATGATTTCCAAATCCT-3'

EMA1-P2

5'-TT GCGGCCGC TTAGTAAAATAGAGTAGAGT-3'

[0012] (2) According to the instructions of the whole blood DNA extraction kit, the whole blood DNA containing the worm strains was extracted, and the extracted whole blood DNA of the worm strains was subjected to routine PCR amplification, and the PCR products were sent to Beijing Dingguo Biotechnology Company for sequencing analysis. The reaction system is 25 μL: 30s / 94°C, 30s / 55°C, 45s / 72°C; 30 cycles. Amplify the target gene fragment of 819bp....

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Abstract

The invention relates to a preparation method of an indirect ELISA detection kit based on theileria equi EMA1. The preparation method comprises the preparation of theileria equi EMA1 and the establishment of a theileria equi disease rELISA detection method. The preparation method overcomes the defects of high cost, poor specificity and time consuming of a conventional detection kit, and establishes the theileria equi disease rELISA detection method which is suitable for the clinical diagnosis and epidemiological monitoring of theileria equi disease.

Description

technical field [0001] The invention relates to gene recombination technology, and aims at the cloning, expression and preparation of the EMA1 gene of Theileria equina and the construction of the rELISA antibody detection kit, especially the preparation method of the indirect ELISA detection kit based on the surface protein 1 of the Theileria equine merozoite. Background technique [0002] Equine theilesisis is a tick-borne blood protozoan disease caused by the equine worm. The disease mainly infects equines (horses, mules, donkeys and zebras) and can be acute, subacute or chronic. It usually presents with fever, anemia, jaundice, hepatosplenomegaly, intravascular hemolysis, visible mucous membrane staining and hemoglobinuria, etc., but the clinical symptoms are not obvious during chronic infection; sick horses will carry worms for life even after later recovery, As a source of infection, it is transmitted to other equine animals; the disease is widely distributed in tropic...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893
Inventor 巴音查汗·盖力克张杨郭庆勇宋瑞其刘世芳李永畅王盼举闻秀秀谢小婉张梦圆
Owner XINJIANG AGRI UNIV
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